【摘要】 通过生物信息学软件分析金黄色葡萄球菌转铁蛋白结合蛋白(StbA)的主要抗原表位区,PCR扩增StbA主要抗原表位区的编码序列;将该序列插入表达载体pGEX-4T-1中,测序验证后转化大肠杆菌BL21(DE3)进行IPTG诱导表达。结果表明,重组菌能表达出分子质量约为54 000u的重组融合蛋白。表达产物经亲和层析纯化后可获得较高纯度的目的蛋白。Western-blot检测证实该重组蛋白能与乳腺炎患病奶牛恢复期血清发生阳性反应。ELISA结果显示,重组蛋白免疫新西兰大白兔能产生高效价的抗血清,提示StbA主要抗原表位区能刺激机体产生强大的免疫应答,StbA是一个有潜力的保护性抗原候选分子。更多还原

【Abstract】 Bioinformatics software was used to analyze the major antigenic epitope region of staphylococcal transferrin-binding protein A(StbA).The encoding sequence was amplified by PCR and cloned into pGEX-4T-1expression vector.After being sequenced,the recombinant plasmid was transformed into Escherichia coli BL21(DE3).The transformed bacteria were induced with IPTG and produced a recombinant protein of 54 ku in molecular mass.Subsequently,the expressed fusion protein was purified by High-Affinity GST Resin.Western-blot confirmed that the purified protein could react with cow serum infected with Staphylococcus aureus.High titer antiserum could be obtained by immunizing New Zealand rabbit with purified recombinant StbA.The result indicated that the major antigenic epitope region of StbA could induce strong immune response in rabbit.In conclusion,StbA is a potential candidate as a protective antigen against S.aureusinfection.更多还原